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Bovine interferon-tau expression in Escherichia coli and identification of its biological activities

Published online by Cambridge University Press:  03 March 2009

Gao Fang-Fang ,
Wu Zhong-Yi  and
Zeng Shen-Ming
Gao Fang-Fang
Affiliation:
College of Animal Science and Technology, China Agricultural University, Beijing 100094, China
Wu Zhong-Yi
Affiliation:
Beijing Research Center of Agro-biotechnology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
Zeng Shen-Ming*
Affiliation:
College of Animal Science and Technology, China Agricultural University, Beijing 100094, China
*
*Corresponding author. E-mail: zengsm@cau.edu.cn
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Abstract

The bovine interferon-tau (bIFN-τ) gene, with signal sequence, was obtained through polymerase chain reaction (PCR) from bovine early embryos and subcloned into a pGEM-T vector. After being verified, the fragments, with or without signal sequence, were inserted into the expression vector pET-30a(+). Two recombinant plasmids were induced to express the recombinant proteins by isopropyl β-d-1-thiogalactopyranoside. The results showed that the bIFN-τ gene could be obtained from five bovine blastocysts by PCR without extraction of genomic DNA. It had 99% homology with nucleotides and 97% with amino acids in the GenBank sequence (accession number: XM_593584). The products of recombinant bIFN-τ, minus signal sequence, expressed in pET-30a(+) were analysed by SDS-PAGE. A new 20 kDa protein was detected and its molecular weight was as expected. The antiviral activity of recombinant bIFN-τ was 1×104 IU/mg using a standard cytopathic reduction assay. Marked morphological changes were induced by recombinant bIFN-τ in bovine endometrial epithelial cells. The cell volume was larger than that of controls and a lot of vesicles appeared in the cytoplasm after 24 h culture in the presence of 2.9 μg/ml recombinant bIFN-τ. In conclusion, a purified recombinant, biologically active bIFN-τ was obtained in this experiment.

Keywords

bovine IFN-τ genecloneexpressionbiological activity
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 3 , December 2008 , pp. 189 - 195
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2008, 16(2): 208–213

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